Comparison of miRNAs and Their Targets in Seed Development between Two Maize Inbred Lines by High-Throughput Sequencing and Degradome Analysis

<div><p>MicroRNAs (miRNAs) play an important role in plant growth, development, and response to environment. For identifying and comparing miRNAs and their targets in seed development between two maize inbred lines (i.e. PH6WC and PH4CV), two sRNAs and two degradome libraries were constructed. Through high-throughput sequencing and miRNA identification, 55 conserved and 24 novel unique miRNA sequences were identified in two sRNA libraries; moreover, through degradome sequencing and analysis, 137 target transcripts corresponding to 38 unique miRNA sequences were identified in two degradome libraries. Subsequently, 16 significantly differentially expressed miRNA sequences were verified by qRT-PCR, in which 9 verified sequences obviously target 30 transcripts mainly involved with regulation in flowering and development in embryo. Therefore, the results suggested that some miRNAs (e.g. miR156, miR171, miR396 and miR444) related reproductive development might differentially express in seed development between the PH6WC and PH4CV maize inbred lines in this present study.</p></div

The Role of the Rho/ROCK Pathway in Ang II and TGF-β1-Induced Atrial Remodeling - Fig 2

<p>The upper panel depicts mRNA expression levels of TGF-β1 (A), CTGF (B), collagen-I (C), RhoA (D) and ROCK-1 (E) in the canine left atrium following 4 weeks of RAP. The middle panel depicts the protein expression levels of TGF-β1 (F), CTGF (G), collagen I (H) and ROCK-1 (I) in the canine left atrium following 4 weeks of RAP. The lower panel depicts the immunohistochemical results pertaining to collagen-I (J), CTGF (K) and ROCK-1 (L) in the hearts (×400). C, control group; P, pacing group. *P<0.05, **P<0.01 vs group C. (n = 6 for Group C, n = 9 for Group P).</p

Summary of miRNA sequencing in the PH6WC and PH4CV libraries.

<p>(a) The mature maize cobs from the PH6WC and PH4CV lines; (b) Overview of sRNA sequences in the PH6WC and PH4CV libraries; (c) Length distributions of clean reads in the PH6WC and PH4CV libraries; (d) miRNA classification in the PH6WC and PH4CV libraries.</p

Differentially expressed miRNAs and their targets involved in seed development between the PH6WC and PH4CV lines.

<p>Differentially expressed miRNAs and their targets involved in seed development between the PH6WC and PH4CV lines.</p

The effect of TGF-β1 on the Smad signal pathway.

<p>(A) TGF-β1 upregulated the expression of Smad2/3. (B) Sim and Losinhibited the protein expression of Smad2/3 in the atrial CFs treated with TGF-β1. (C) The ratio of GTP-RhoA/total RhoA following different treatments. *P<0.05, **P<0.01 vs TGF-β1 group, #<i>P</i><0.05, ##<i>P</i><0.01 vs Y27632+TGF-β1 group. (n = 5 for each group).</p

qRT-PCR validated miRNAs and their targets in the PH6WC and PH4CV lines.

<p>qRT-PCR validated miRNAs and their targets in the PH6WC and PH4CV lines.</p

Comparison of hydroxyproline expression between the two groups.

<p>LA, left atrium; RA, right atrium; LV, left ventricle; RV, right ventricle. *P<0.05, **P<0.01. (n = 6 for Group C, n = 9 for Group P).</p

Representative HE staining (up panel) and Masson’s trichrome staining (down panel) of the atrial myocardium.

<p>(A) Structurally normal atrial myocardium in the control group and severe fibrosis in the pacing group (×100). (B) Atrial myocardium with HE staining under ×400 light microscopy. (C) A small amount of collagen is visible within the atrial myocardium of the control group, and an increased amount of collagen is visible within the atrial myocardium following pacing (×100). The red areas represent myocytes, and the blue areas represent collagen. (D) Atrial myocardium with Masson’s trichrome staining under ×400 light microscopy. C, control group; P, pacing group. (n = 6 for Group C, n = 9 for Group P).</p

The effect of the Ang II pathway on atrial CF proliferation (upper panel) and the effect of Ang II on the mRNA expression of the AF factors in the atrial CFs (lower panel).

<p>(A) The effects of different concentrations of Ang II on the proliferation of the atrial CFs. (B, C, D) The effects of different concentration of Los (B), Sim (C) and Y27632 (D) on Ang II induced cell proliferation. Ang II upregulated the expression of ROCK-1 (E), CTGF (F) and TGF-β1 (G). Sim, Los and Y27632 downregulated the Ang II induced expression of TGF-β1 (J), CTGF (I) and ROCK-1 (H). *P<0.05, **P<0.01 vs Group C, #<i>P</i><0.05, ##<i>P</i><0.01 vs Group Ang II.(n = 5 for each group).</p

qRT-PCR validation of unique miRNA sequences in the PH6WC and PH4CV lines.

<p>qRT-PCR validation of unique miRNA sequences in the PH6WC and PH4CV lines.</p